期刊
JOURNAL OF CONTROLLED RELEASE
卷 228, 期 -, 页码 1-8出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jconrel.2016.01.032
关键词
Small-interfering RNA; Polycation liposomes; Freeze-thaw; siRNA encapsulation
资金
- Japan Society for the Promotion of Science (JSPS)
- Kurata Memorial Hitachi Science and Technology Foundation
Small interfering RNA (siRNA) has the potential to be a candidate as a cure for intractable diseases. However, an appropriate vector is required for siRNA delivery because of the low transfection efficiency of siRNA without a vector and its easy degradation in vivo. Here, we report a simple, only one step, and efficient method for siRNA encapsulation into a lipidic nanocarrier by freeze-thawing: siRNA was entrapped between the lipid layers of multi-layer liposomes by freeze-thawing of lipoplexes composed of polycation liposomes (PCLs) and siRNA. siRNA-holding capacity to the PCL was increased by repeating freeze-thaw of the lipoplex up to 5 cycles. Although siRNA in the conventional lipoplex was degraded after incubation in 90% fetal bovine serum for 72 h, siRNA in the frozen and thawed lipoplex was not degraded. Interestingly, we found that the lipoplex formed a packed multi-layer structure after the freeze-thawing of single-layer PCL and siRNA complex, suggesting that siRNA exists between the lipid layers working as a binder. The frozen and thawed lipoplex showed significantly higher knockdown efficacy compared with the conventional lipoplex. In addition, PEGylated freeze-thawed lipoplexes delivered a higher amount of siRNA to a tumor in vivo compared with the PEGylated conventional ones. These results provide an attractive strategy for one-step encapsulation of siRNA into liposomes by freeze-thawing. (C) 2016 Elsevier B.V. All rights reserved.
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